Identification of Campylobacter jejuni promoter sequences.
A promoterless lacZ shuttle vector, which allowed screening of promoters by beta-galactosidase activity in Campylobacter jejuni and Escherichia coli, was developed. Chromosomal DNA fragments from C. jejuni were cloned into this vector; 125 of 1,824 clones displayed promoter activity in C. jejuni. Eleven clones with strong promoter activity in C. jejuni were further characterized. Their nucleotide sequences were determined, and the transcriptional start sites of the putative promoters in C. jejuni were determined by primer extension. Only 6 of these 11 promoters were functional in E. coli. The 11 newly characterized and 10 previously characterized C. jejuni promoters were used to establish a consensus sequence for C. jejuni promoters. The 21 promoters were found to be very similar. They contain three conserved regions, located approximately 10, 16, and 35 bp upstream of the transcriptional start point. The -10 region resembles that of a typical sigma70 E. coli promoter, but the -35 region is completely different. In addition a -16 region typical for gram-positive bacteria was identified.
Wösten
M M
MM
Department of Bacteriology, Institute of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Universiteit Utrecht, The Netherlands.
Boeve
M
M
Koot
M G
MG
van Nuenen
A C
AC
van der Zeijst
B A
BA
D001483:Base Sequence; D001665:Binding Sites; D016123:Campylobacter jejuni / Q000235:genetics*; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004269:DNA, Bacterial / Q000235:genetics*; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D015723:Gene Library; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010442:Peptide Chain Initiation, Translational; D011401:Promoter Regions, Genetic; D012329:RNA, Bacterial; D012333:RNA, Messenger; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D012808:Sigma Factor / Q000235:genetics; D014158:Transcription, Genetic
D004269:DNA, Bacterial; D012329:RNA, Bacterial; D012333:RNA, Messenger; D012808:Sigma Factor; C081775:RNA polymerase sigma 70; D012321:DNA-Directed RNA Polymerases